Anti-DYKDDDDK Magarose beads
洋甘菊提取液MSDS英文版
1. IDENTIFICATION OF THE SUBSTANCE/TREPARATION AND THE COMPANY/UNDERTAKING3.HAZARDS IDENTIFICATION4. FIRST AID MEASURESMATERIAL SAFETY DATA SHEETProduct name:Supplier:Tel:EMERGENCY OVERVIEW: May cause skin irritation and/or dermatitisPrinciple routes of exposure: Inhalation: Ingestion: Skin contact: Eye contact:SkinMay cause irritation of respiratory tract May be harmful if swallowed May cause allergic skin reaction Avoid contact with eyesStatements of hazard MAY CAUSE ALLERGIC SKIN REACTION.Statements of Spill of Leak Label Eliminate all ignition sources. Absorb and/or contain spill with inert materials (e.g., sand, vermiculite). Then place in appropriate container. For large spills, use water spray to disperse vapors, flush spill area. Prevent runoff from entering waterways or sewers.General advice:POSITION/INFORMATION ON INGREDIENTSInhalation:Skin contact:Ingestion:Eye contact:Protection of first – aiders:Medical conditions aggravated by exposure: In the case of accident or if you fell unwell, seek medical advice immediately (show the label where possible).Move to fresh air, call a physician immediately.Rinse immediately with plenty of water and seek medical adviceDo not induce vomiting without medical advice.In the case of contact with eyes, rinse immediately with plenty of water and seek medical advice.No information availableNone knownSuitable extinguishing media:Specific hazards:Special protective equipment for firefighters:Flash point:Autoignition temperature:NFPA rating Use dry chemical, CO2, water spray or “alcohol” foam Burning produces irritant fumes.As in any fire, wear self-contained breathing apparatus pressure-demand, MSHA/NIOSH (approved or equivalent) and full protective gearNot determinedNot determinedNFPA Health: 1 NFPA Flammability: 1 NFPA Reactivity: 0Personal precautions: Environmental precautions: Methods for cleaning up: Use personal protective equipment.Prevent product from entering drains.Sweep up and shovel into suitable containers for disposalStorage:7. HANDLING AND STORAGE5.FIRE-FIGHTING MEASURES6. ACCIDENTAL RELEASE MEASURESRoom temperature Handling:Safe handling advice: Incompatible products:Use only in area provided with appropriate exhaust ventilation.Wear personal protective equipment.Oxidising and spontaneously flammable productsEngineering measures: Respiratory protection: Skin and body protection:Eye protection: Hand protection: Hygiene measures:Ensure adequate ventilation.Breathing apparatus only if aerosol or dust is formed. Usual safety precautions while handling the product will provide adequate protection against this potential effect. Safety glasses with side-shieldsPVC or other plastic material glovesHandle in accordance with good industrial hygiene and safety practice.Melting point/range: Boiling point/range: Density: Vapor pressure: Evaporation rate: Vapor density: Solubility (in water): Flash point:Autoignition temperature:No Data available at this time. No Data available at this time. No data available No data available No data available No data available No data available Not determined Not determinedStability: Stable under recommended storage conditions. Polymerization: None under normal processing.Hazardous decomposition products: Thermal decomposition can lead to release of irritating gases and vapours such as carbon oxides.Materials to avoid: Strong oxidising agents.10. STABILITY AND REACTIVITY9. PHYSICAL AND CHEMICAL PROPERTIES8. EXPOSURE CONTROLS/PERSONAL PROTECTION11. TOXICOLOGICAL INFORMATIONConditions to avoid: Exposure to air or moisture over prolonged periods.Product information Acute toxicityChronic toxicity:Local effects: Chronic exposure may cause nausea and vomiting, higher exposure causes unconsciousness.Symptoms of overexposure may be headache, dizziness, tiredness, nausea and vomiting.Specific effects:May include moderate to severe erythema (redness) and moderate edema (raised skin), nausea, vomiting,headache.Primary irritation: Carcingenic effects: Mutagenic effects: Reproductive toxicity:No data is available on the product itself. No data is available on the product itself. No data is available on the product itself. No data is available on the product itself.Mobility:Bioaccumulation: Ecotoxicity effects: Aquatic toxicity:No data available No data available No data availableMay cause long-term adverse effects in the aquatic environment.12. ECOLOGICAL INFORMATION13. DISPOSAL CONSIDERATIONSWaste from residues/unused products:Contaminated packaging:Waste disposal must be in accordance with appropriate Federal, State and local regulations. This product, if unaltered by use, may be disposed of treatment at a permitted facility or as advised by your local hazardous waste regulatory authority. Residue from fires extinguished with this material may be hazardous.Do not re-use empty containers.UN/Id No:Not regulated14. TRANSPORT INFFORMATIONDOTProper shipping name: Not regulatedTGD(Canada)WHMIS hazard class: Non - controlledIMDG/IMOIMDG – Hazard Classifications Not ApplicableIMO – labels:15. REGULATORY INFOTMATION International Inventories16. OTHER INFORMATIONPrepared by: Health & SafetyDisclaimer: The information and recommendations contained herein are based upon tests believed to be reliable.However, XABC does not guarantee the accuracy or completeness NOR SHALL ANY OF THIS INFORMATION CONSTITUTE A WARRANTY, WHETHER EXPRESSED OR IMPLIED, AS TO THE SAFETY OF THE GOOD, THE MERCHANTABILITY OF THE GOODS, OR THE FITNESS OF THE FITNESS OF THE GOODS FOR A PARTICULAR PURPOSE. Adjustment to conform to actual conditions of usage maybe required. XABC assumes no responsibility for results obtained or for incidental or consequential damages, including lost profits arising from the use of these data. No warranty against infringement of any patent, copyright or trademark is made or implied.End of safety data sheet。
常见蛋白质标签总结
常见蛋白质标签总结（Flag、HA、cMyc、CBP等）Protein tags are peptide sequences genetically grafted onto a recombinant protein. Often these tags are removable by chemical agents or by enzymatic means, such as proteolysis or intein splicing. Tags are attached to proteins for various purposes.一、氨基酸标签（含小肽标签）A stretch of amino acids is added to the protein and enables the recovery of the labelled protein by its unique affinity. Usually its easiest to add the tag to either end of the protein to ensure its accessibility and not to disturb the protein folding.1.组氨酸标签（His tag）一般为6个组氨酸，用Ni2+（Cu2+）亲和层析纯化2.FLAG tag ：N-DYKDDDDK-C ，recovered with specific antibody3.HA tag： an epitope derived from the Influenza protein haemagglutinin(HA，禽流感病毒血凝素)，e.g. N-YPYDVPDYA-C，recovery with anHA antibody4.MYC tag： an epitope derived from the human proto-oncoprotein MYC，e.g.N-ILKKATAYIL-C, N-EQKLISEEDL-C，recovery with an MYCantibody5.SBP tag：Streptavidin Binding Peptide，链霉亲合素结合肽，38 amino acidtag (MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP)，更多参考在Sigma6.CBP tag：钙调蛋白结合肽（CBP; 26aa）钙调蛋白结合肽与钙调素结合是Ca2+依赖的，这种结合不受标签所处的位置影响（N端和C端均可），在中性pH条件下使用2mM EGTA可以很方便的将目标蛋白洗脱下来。
常见蛋白质标签总结
/bbs/home.php?mod=space&uid =34800&do=blog&id=38530常见蛋白质标签总结（Flag、HA、cMyc、CBP等）Protein tags are peptide sequences genetically grafted onto a recombinant protein. Often these tags are removable by chemical agents or by enzymatic means, such as proteolysis or intein splicing. Tags are attached to proteins for various purposes.一、氨基酸标签（含小肽标签）A stretch of amino acids is added to the protein and enables the recovery of the labelled protein by its unique affinity. Usually its easiest to add the tag to either end of the protein to ensure its accessibility and not to disturb the protein folding.1.组氨酸标签（His tag）一般为6个组氨酸，用Ni2+（Cu2+）亲和层析纯化2.FLAG tag ：N-DYKDDDDK-C ，recovered with specific antibody3.HA tag： an epitope derived from the Influenza protein haemagglutinin (HA，禽流感病毒血凝素)，e.g. N-YPYDVPDYA-C，recovery with an HAantibody4.MYC tag： an epitope derived from the human proto-oncoprotein MYC，e.g.N-ILKKATAYIL-C, N-EQKLISEEDL-C，recovery with an MYCantibody5.SBP tag：Streptavidin Binding Peptide，链霉亲合素结合肽，38 amino acidtag (MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP)，更多参考在Sigma6.CBP tag：钙调蛋白结合肽（CBP; 26aa）钙调蛋白结合肽与钙调素结合是Ca2+依赖的，这种结合不受标签所处的位置影响（N端和C端均可），在中性pH条件下使用2mM EGTA可以很方便的将目标蛋白洗脱下来。
兰嘉丝汀
【兰嘉丝汀幻彩亮彩去角质面膜（橘色胶囊）】特别添加柳橙、柠檬、葡萄柚及杏桃核颗粒。
能有效使暗沉粗糙的肌肤，立即恢复柔嫩透亮，更好吸收保养品。
所有肤质适用。
使用方法：将脸打湿后，避开眼周，涂上薄薄一层，用指腹轻轻画圈按摩。
最后以温水冲洗干净。
一个胶囊的使用量为一次。
【兰嘉丝汀幻彩珍珠活力面膜（黄色胶囊）】（右一）特含有莲花及维他命A，可立即补充新鲜美丽能量。
针对疲累、压力型肌肤，可立即醒肤，恢复珍珠般明亮光采，适用于所有肤质。
使用方法：厚敷于全脸及颈部，停留5分钟，用面纸将多余的面膜轻轻擦拭。
一个胶囊的使用量为一次。
【兰嘉丝汀理肤银杏隔离霜SPF50/PA+++ 3ML】（左二)运用独家防晒技术，完美延展科技，全面覆盖肌肤，不堵塞毛孔，不生成粉刺，在肌肤形成透明保护膜，高效抵御各种肌肤伤害源。
使用方法：取适量用于面霜之后，均匀涂抹与面部，避开眼部。
出门前20分钟使用。
【兰嘉丝汀水润身体防晒液SPF30 30ml 】（左一）双重防护，高效预防UV A/UVB，专利天然RPF复合因子，可同时中和95%因外在环境所产生的自由基。
专为亚洲肌肤需求设计，防止肌肤晒黑，运用特殊水润配方。
使用方法：于外出前30分钟涂于身体暴露部位，每过一段时间加涂一次新七白美白嫩肤面膜洁肤后，取适量（两粒葡萄大小，约6g），轻柔均匀地涂抹于面部，停留10-15分钟，再以温水洗净，每周使用3-4次。
搭配佰草集新七白美白系列产品共同使用，效果尤佳。
清肌养颜太极泥先清：首先使用黑泥，清洁双手并擦干，用面膜调棒取适量黑泥，涂抹薄薄一层于面部，至掩盖肤色铺展均匀即可，注意避开眼部周围。
加轻轻按摩以，使黑泥中的本草成分渗透并作用于肌肤。
约5分钟后，用洁面棉擦去黑泥，再用温水冲洗干净；后补：接着使用白泥，黑泥洗去后，在面部半湿状态下，用面膜调棒取适量白泥，均匀涂抹于面部；用食指和中指以划圈的方式轻柔按摩面部2-3分钟，按摩后，再使白泥在脸部停留8分钟左右，使肌肤充分吸收营养成分。
FlowComp
Kit ContentsFlowComp ™ Dynabeads ® contains ~1 × 109 (~10 mg) beads/mL in phosphate buffered saline (PBS), pH 7.4, with 0.1% bovine serum albumin (BSA) and 0.02% sodium azide as a preservative. FlowComp ™ Human CD3 Antibody contains monoclonal CD3 antibody in PBS with 0.5% BSA and 0.02% sodium azide. FlowComp ™ Release Buffer contains modified biotin in 0.1% BSA and 2 mM EDTA.Caution: Sodium azide may react with lead and copper plumbing to form highly explosive metal azides.Product DescriptionFlowComp ™ Human CD3 is intended for positive magnetic isolation of CD3+ T cells from human peripheral bloodmononuclear cells (PBMCs). The isolated cells are highly pure, viable, and bead-free (fig. 1). In the first step, FlowComp ™ Human CD3 Antibody is added and binds to the target cells. In the second step, CD3+ T cells, that have bound the specific antibodies, are captured by the FlowComp ™ Dynabeads ®. In the third and last step, the cells are released from the FlowComp ™ Dynabeads ®.Dynabeads ® FlowComp ™ Human CD3Isolation from PBMCFor research use only. Not for human or animal therapeutic or diagnostic use.Catalog no. 11365D Store at 2˚C to 8˚CRev. Date: February 2012 (Rev. 001)Downstream ApplicationsIsolated cells are bead-free and may be used directly in anydownstream application including flow cytometry. The cells readily proliferate in response toDynabeads ® Human T Activator CD3/CD28 and can be measured by incorporating EdU or in a CFSE assay.Required Materials• Magnet (DynaMag ™ portfolio).See /magnets for recommendations.• Mixer allowing tiltingand rotation of tubes (e.g. HulaMixer ® Sample Mixer).• Isolation Buffer:Ca 2+ and Mg 2+ free PBSsupplemented with 0.1% BSA and 2 mM EDTA.Note: BSA can be replaced by human serum albumin (HSA) or 2% FBS/FCS.• Optional: Flow cytometry antibodies. We recommend using anti-CD3clone UCHT-1 from Caltag Medsystems as primary fluorescent antibody for flow staining of cells after isolation. Optional clones: OKT3, HITa3.• Optional: For viability analysis, SYTOX ® Red is recommended.General Guidelines• Visit /cellisolation and follow ourQuickLinks for recommended sample preparation procedures.• Use a mixer that provides tilting and rotation of the tubes to ensure thatbeads do not settle in the tube.• This product should not be used with the MPC ™-1 magnet(Cat. no. 12001D).• Avoid air bubbles (foaming) during pipetting.• Never use less than the recommended volume of beads.• Carefully follow the recommended pipetting volumes and incubationtimes.• Keep all buffers cold.• To avoid unspecific labeling of cells during flow staining, werecommend using gammaglobulin prior to staining with primary fluorescent antibody.• For better purity, repeat the washing step once or transfer the bead-bound cells to a new tube before adding the FlowComp ™ Release Buffer.• All incubations at room temperature can also be performed at2°C to 8°C.ProtocolThis protocol is intended for isolation of bead-free CD14+ monocytes, starting with 5 × 107 PBMC/test where typically 10–20% are CD14+ monocytes. When working with higher cell numbers/number of tests, scale up all volumes accordingly, as shown in Table 1.Wash the BeadsSee Table 1 for volume recommendations.1. Resuspend the beads in the vial (i.e. vortex for >30 sec, or tilt and rotatefor 5 min).2. Transfer the desired volume of beads to a tube.3. Add the same volume of Isolation Buffer, or at least 1 mL, andresuspend.4. Place the tube in a magnet for 1 min and discard the supernatant.5. Remove the tube from the magnet and resuspend the washed beadsin the same volume of Isolation Buffer as the initial volume of beads (step 2).Prepare Cells• Prepare a PBMC suspension according to “General Guidelines”.Resuspend the cells at 1 × 108 cells/mL in Isolation Buffer. • Prepare approximately 10 mL of Isolation Buffer per 5 × 107cells.PBMC: ~2 × 109Figure 1: Purity of human CD3+ cells isolated from PBMC using Dynabeads ® FlowComp ™Human CD3.For support visit /support or *****************************©2012 Life Technologies Corporation. All rights reserved. The trademarks mentioned herein are the property of Life Technologies Corporation or their respective owners, except where otherwise stated. LIFE TECHNOLOGIES AND/OR ITS AFFILIATE(S) DISCLAIM ALL WARRANTIES WITH RESPECT TO THIS DOCUMENT, EXPRESSED OR IMPLIED, INCLUDING BUT NOT LIMITED TO THOSE OF MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE. IN NO EVENT SHALL LIFE TECHNOLOGIES AND/OR ITS AFFILIATE(S) BE LIABLE, WHETHER IN CONTRACT, TORT, WARRANTY, OR UNDER ANY STATUTE OR ON ANY OTHER BASIS FOR SPECIAL, INCIDENTAL, INDIRECT, PUNITIVE, MULTIPLE OR CONSEQUENTIAL DAMAGES IN CONNECTION WITH OR ARISING FROM THIS DOCUMENT, INCLUDING BUT NOT LIMITED TO THE USE THEREOF .SPEC-06682on labels is the symbol for catalog number.REF Table 1: Volumes for human CD3+ T cells. This protocol is scalable from 1 × 107 to 5 × 108 PBMC. larger tube than recommended in step 14 to successfully remove the biotin in the sample.** When incubating, tilt and rotate the vial so the cells and beads are kept in the bottom of the tube. Do not perform end-over-end mixing if the volume is small relative to the tube size.Description of MaterialsFlowComp ™ Dynabeads ® are uniform, superparamagnetic polystyrene beads (2.8 μm in diameter) coated with modified streptavidin. FlowComp ™ Human CD3 Antibody contains a DSB-X conjugated monoclonal mouse anti-human CD3. FlowComp ™ Release Buffer contains a modified biotin that displaces the modified biotin on the antibody to release cells from the beads.Limited Use Label LicenseThe purchase of this product conveys to the purchaser the limited, nontransferable right to use the purchased amount of the product only to perform internal research for the sole benefit of the purchaser. No right to resell this product or any of itscomponents is conveyed expressly, by implication, or by estoppel. This product is for internal research purposes only and is not for use in commercial applications of any kind, including, without limitation, quality control and commercial services such as reporting the results of purchaser’s activities for a fee or other form of consideration. For information on obtaining additional rights, please contact outlicensing@ or Out Licensing, Life Technologies, 5791 Van Allen Way, Carlsbad, California 92008.Manufactured by Life Technologies AS, Norway. Life Technologies AS complies with the Quality System Standards ISO 9001:2008 and ISO 13485:2003.Limited Product WarrantyLife Technologies Corporation and/or its affiliate(s) warrant their products as set forth in the Life Technologies' General Terms and Conditions of Sale found on Life Technologies' website at /termsandconditions . If you have any questions, please contact Life Technologies at /support .Isolate CellsThis protocol is based on 5 × 107PBMC, but is directly scalable from 1 × 107to 5 × 108 cells, according to Table 1.1. Transfer 500 μL (5 × 107 cells) prepared cells to a tube and add 25 μLFlowComp ™ Human CD3 antibody. 2. Mix well and incubate for 10 min at 2°C to 8°C.3. Wash by adding 2 mL Isolation Buffer and centrifuge for 8 min at350 × g.4. Remove the supernatant and resuspend in 1 mL Isolation Buffer.5. Add 75 μL washed FlowComp ™ Dynabeads ® and mix well(e.g. vortex 2–3 seconds).6. Incubate for 15 min at room temperature under rolling and tilting.7. Add 1 mL isolation buffer, pipet 2–3 times (or vortex 2–3 seconds) andplace the tube in a magnet for 2 min.8. While the tube is still in the magnet, carefully remove and discard thesupernatant containing the CD3 negative cells.9. Repeat steps 7–8 to wash the bead-bound CD3+ cells. These steps arecritical to obtain a high purity of isolated cells.Release Cells10. Resuspend the bead-bound cells in 1 mL Release Buffer.11. Incubate for 10 min with rolling and tilting at room temperature.12. Pipet 10 times to efficiently release the cells and place in a magnet for2 min. Avoid foaming.13. Transfer the supernatant containing the bead-free CD3+ cells to a newtube, and again place on the magnet for 1 min to remove any residual beads. Transfer again the supernatant containing the bead-free cells to a new tube.14. Add 2 mL Isolation Buffer followed by centrifugation for 8 min at350 × g. Discard the supernatant and resuspend the cell pellet in preferred medium.Keep the cells on 2°C to 8°C until further use in downstream applications.Related Products。
Halo-Trap Agarose 产品说明书
The ChromoTek Halo-Trap Agarose consists of an anti-Halo-tag Nanobody (VHH), which is covalently bound to agarose beads. Halo-Trap Agarose is used to immunoprecipitate Halo-tagged fusion proteins from cell extracts of various organisms like mammals, plants, bacteria, yeast, insects etc. in the presence or absence of a covalently bound ligand. The interaction between Halo-Trap and the Halo-tagged fusion protein is reversible.Ligand: Anti-Halo-tag single domain antibody fragment (VHH, Nanobody)Reactivity: Specifically binds to Halo-tag (modified variant of the bacterial haloalkane dehalogenase enzyme from Rhodococcus rhodochrous) in the absence or presence of covalently bound chloralkane-based ligands. Binding capacity: 7.5-10 µg of recombinant Halo-tag per 25 µL bead slurryBead size: 90 µm (cross-linked 4 % agarose beads)Buffer compatibility: See Wash buffer compatibility table.Storage buffer: 20 % ethanolStorage conditions: Upon receipt store at +4°C. Do not freeze!Stability: Stable for 1 year upon receipt.Shipment: Shipped at ambient temperature.RRID: AB_2827595Required buffer solutionsNEW: Update of Wash buffer components.Buffer CompositionLysis buffer10 mM Tris/Cl pH 7.5, 150 mM NaCl, 0.5 mM EDTA, 0.5 % Nonidet™ P40 Substitute (adjust the pH at +4°C)RIPA buffer10 mM Tris/Cl pH 7.5, 150 mM NaCl, 0.5 mM EDTA, 0.1 % SDS, 1 % Triton™ X-100, 1 %deoxycholate (adjust the pH at +4°C)Dilution buffer10 mM Tris/Cl pH 7.5, 150 mM NaCl, 0.5 mM EDTA (adjust the pH at +4°C)Wash buffer10 mM Tris/Cl pH 7.5, 150 mM NaCl, 0.05 % Nonidet™ P40 Substitute, 0.5 mM EDTA (adjust the pH at +4°C)2x SDS-sample buffer120 mM Tris/Cl pH 6.8, 20 % glycerol, 4 % SDS, 0.04 % bromophenol blue, 10 % β-mercaptoethanolAcidic elution buffer 200 mM glycine pH 2.5 or 100 mM citric acid pH 3.0 (adjust the pH at +4°C)Neutralization buffer 1 M Tris pH 10.4 (adjust the pH at +4°C)Note: Use your equivalent cell lysis buffer for other cell types like yeast, plants, insects, bacteria.Note: Consider using a Wash buffer without detergent for co-immunoprecipitation.Buffer ingredients Max. concentrationDTT10 mMNaCl 1 MNonidet™ P40 Substitute tested up to 2 %SDS0 %Triton™ X-100tested up to 1 %Urea 4 MProduct Product code SizeHalo-Trap Agarose ota-1010 reactions (250 µL slurry)ota-2020 reactions (500 µL slurry)ota-100100 reactions (2.5 mL slurry)ota-200200 reactions (5 mL slurry)ota-400400 reactions (10 mL slurry)Halo-Trap Agarose Kit otak-2020 reactions (500 µL slurry) including buffersCell materialThe following protocol describes the preparation of mammalian cell lysate!For other type of cells, we recommend using 500 µg of cell extract and start the protocol with step Bead equilibration.Mammalian cell lysisNote: Harvesting of cells and cell lysis should be performed with ice-cold buffers. We strongly recommend to add protease inhibitors to the Lysis buffer to prevent degradation of your target protein and its binding partners.For one immunoprecipitation reaction, we recommend using ~106-107 cells.1.Choice of lysis buffer:·For cytoplasmic proteins, resuspend the cell pellet in 200 µL ice-cold Lysis buffer by pipetting up and down. Supplement Lysis buffer with protease inhibitor cocktail and 1 mM PMSF (not included).·For nuclear/chromatin proteins, resuspend cell pellet in 200 µL ice-cold RIPA buffer supplemented(f.c. 2.5 mM), protease inhibitor cocktail and PMSF (f.c. 1with DNaseI (f.c. 75-150 Kunitz U/mL), MgCl2mM) (not included).2.Place the tube on ice for 30 min and extensively pipette the suspension every 10 min.3.Centrifuge cell lysate at 17,000x g for 10 min at +4°C. Transfer cleared lysate (supernatant) to a pre-cooled tube and add 300 µL Dilution buffer supplemented with 1 mM PMSF and protease inhibitor cocktail (not included). If required, save 50 µL of diluted lysate for further analysis (input fraction).Bead equilibration1.Resuspend the beads by gently pipetting up and down or by inverting the tube. Do not vortex the beads!2.Transfer 25 µL of bead slurry into a 1.5 mL reaction tube.3.Add 500 µL ice-cold Dilution buffer.4.Sediment the beads by centrifugation at 2,500x g for 5 min at +4°C. Discard the supernatant.Note: Alternatively, Spin columns (sct-10; -20; -50) can be used to equilibrate the beads.Protein binding1.Add diluted lysate to the equilibrated beads.2.Rotate end-over-end for 1 hour at +4°C.Washing1.Sediment the beads by centrifugation at 2,500x g for 5 min at +4°C.2.If required, save 50 µL of supernatant for further analysis (flow-through/non-bound fraction).3.Discard remaining supernatant.4.Resuspend beads in 500 µL Wash buffer.5.Sediment the beads by centrifugation at 2,500x g for 5 min at +4°C. Discard remaining supernatant.6.Repeat this step at least twice.7.During the last washing step, transfer the beads to a new tube.Optional: To increase stringency of the Wash buffer, test various salt concentrations e.g. 150-500 mM, and/or add a non-ionic detergent e.g. Triton™ X-100 (see Wash buffer compatibility table for maximal concentrations). Note: Alternatively, Spin columns (sct-10; -20; -50) can be used to wash the beads.Elution with 2x SDS-sample buffer (Laemmli)1.Remove the remaining supernatant.2.Resuspend beads in 80 µL 2x SDS-sample buffer.3.Boil beads for 5 min at +95°C to dissociate immunocomplexes from beads.4.Sediment the beads by centrifugation at 2,500x g for 2 min at +4°C.5.Analyze the supernatant in SDS-PAGE / Western Blot.Note: For Western blot detection we recommend Halo antibody [28A8] (28a8-20; -100).Elution with Acidic elution buffer1.Remove the remaining supernatant.2.Add 50-100 µL Acidic elution buffer and constantly pipette up and down for 30-60 sec at +4°C or roomtemperature.3.Sediment the beads by centrifugation at 2,500x g for 2 min at +4°C.4.Transfer the supernatant to a new tube.5.Immediately neutralize the eluate fraction with 5-10 µL Neutralization buffer.6.Repeat this step at least once to increase elution efficiency.Note: Elution at room temperature is more efficient than elution at +4°C. Prewarm buffers for elution at room temperature.Note: Alternatively, Spin columns (sct-10; -20; -50) can be used to separate the beads.Halo-tag toolbox Product code Halo-Trap Agarose ota-10; -20; -100 Halo-Trap Agarose Kit otak-20Binding Control Agarose bab-20Spin columns sct-10; sct-20; sct-50 Halo VHH, recombinant binding protein ot-250Halo antibody [28A8]28a8-20; -100For product details, information, and ordering visit .*********************ChromoTek GmbHAm Klopferspitz 1982152 Planegg-Martinsried Germanyphone: +49 89 124 148 80 fax: +49 89 124 148 811ChromoTek Inc.62-64 Enter Lane Islandia, NY 11749 USAphone: 631 501 1058 fax: 631 501 1060Only for research applications, not for diagnostic or therapeutic use!ChromoTek and GFP-Trap, RFP-Trap, Myc-Trap, Spot-Trap, Spot-Tag, Spot-Label, Spot-Cap, Nano-Secondary, F2H Kit, and Chromobody are registered trademarks of ChromoTek GmbH, part of Proteintech group. Nano-CaptureLigand and V5-Trap are trademarks of ChromoTek GmbH, part of Proteintech group. Nanobody is a registered trademark of Ablynx, a Sanofi company. Alexa Fluor is a registered trademark of Life Technologies Corporation, a part of Thermo Fisher Scientific Inc. Dynabeads is a trademark of Life Technologies AS, a part of Thermo Fisher Scientific Inc. SNAP-tag is a registered trademark and CLIP-tag is a trademark of New England Biolabs, Inc. Octet is a registered trademark of FortéBio, a Sartorius brand. Other suppliers’ products may be trademarks or registered trademarks of the corresponding supplier each. Statements on other suppliers’ products are given according to our best knowledge.。
Protein A G MagBeads 技术手册说明书
Protein A/G MagBeads Cat. No. L00277Technical Manual No. TM0249 Version 08212013 Index1.Product Description2. Instruction For Use3.Troubleshooting4. General Information1.Product Description1.1Intended UseGenScript Protein A/G MagBeads are ideal for small‐scale antibody purification and immunoprecipitation (IP) of proteins, protein complexes or other antigens.1.2PrincipleThe sample containing antibody is added to the Protein A/G MagBeads. The antibody will bind to beads during a short incubation. Then the beads‐bound antibody may be eluted from the beads by using a magnetic separation rack, or used for immunoprecipitation (IP). A cross‐linking procedure may be needed before IP to prevent co‐elution of the primary antibody. Magnetic separation eliminates the changes of micro tubes, minimizes the loss of sample and removes excessive steps of traditional centrifugation method.1.3Description of MaterialMaterial SuppliedGenScript Protein A/G MagBeads are super paramagnetic beads of average 40 μm in diameter, covalently coated with recombinant Protein A/G. The beads are supplied as 25% slurry in phosphate buffered saline (PBS), pH 7.4, containing 20% ethanol. The Protein A/G MagBeads have a binding capacity of more than 10 mg Goat IgG per 1 ml settled beads (e.g. 4 ml 25% slurry).Protein A/G is a genetically engineered protein (MW≈43 kDa) that combines the IgG binding sites of both Protein A and Protein G. 6×His‐tag was attached to its N‐terminal to facilitate the purification. The secreted Protein A/G contains four Fc‐binding domains from Protein A and two from Protein G, making it a more universal tool to bind and purify immunoglobulins.Cat. No. L00277 Size: 2 ml.Additional Material RequiredMixing/Rotation DeviceMagnetic Separation RackTest tubes and pipettesBuffers and solutions (see below)Additional Buffers NeededBinding/Wash Buffer: 20 mM Na2HPO4, 0.15 M NaCl, pH 7.0Elution Buffer: 0.1 M glycine, pH 2‐3Neutralization Buffer: 1 M Tris, pH 8.51×SDS Sample Buffer: 62.5 mM Tris‐HCl (pH 6.8 at 25°C), 2% w/v SDS, 10% glycerol, 50 mM DTT,0.01% w/v bromophenol blue2.Instruction For UseThe protocol uses 100 μl Protein A/G MagBeads, this may be scaled up or down accordingly.2.1Preparation of the MagBeadspletely resuspend the beads by shaking or vortexing the vial.2.Transfer 100 μl beads into a clean tube.3.Place the tube on a magnetic separation rack to collect the beads. Remove and discard the supernatant.4.Add 1 ml Binding/Wash Buffer to the tube and invert the tube several times to mix. Use the magnetic separationrack to collect the beads and discard the supernatant. Repeat this step twice.2.2Separation of Target IgG1.Resuspend the beads in 100 μl Binding/Wash Buffer.2.Add the sample containing target IgG to the tube and gently invert the tube to mix.3.Incubate the tube at room temperature with mixing (on a shaker or rotator) for 30 – 60 minutes.e the magnetic separation rack to collect the beads and discard the supernatant. If necessary, keep thesupernatant for analysis.5.Add 1 ml Binding/Wash Buffer to the tube and mix well, use the magnetic separation rack to collect the beads anddiscard the supernatant. Repeat the wash step three more times.6.Proceed to elution of isolated IgG (Section 2.3).2.3Elution of Isolated IgG1.Add 100 μl Elution Buffer to the tube and mix well. Incubate for five minutes at room temperature with occasionalmixing.e the magnetic separation rack to collect the beads and transfer the supernatant that contains the eluted IgG intoa clean tube.3.Repeat Step 1 and 2 twice.4.Add 10 μl of Neutralization Buffer to each 100 μl eluate to neutralize the pH. If needed, perform a buffer exchangeby dialysis or desalting.2.4ImmunoprecipitationBound IgG will be co‐eluted along with the target when using elution methods. If the presence of IgG does not disturb desired detection system, go directly to section 2.4.2 below. For applications where co‐elution of the IgG is not desired, the primary IgG can be cross‐linked to the Protein A/G MagBeads as described in section 2.4.1 below.2.4.1Cross‐linking IgG to the Beads1.Add 1 ml 0.2 M triethanolamine, pH 8.2 to the Protein A/G MagBeads with immobilised IgG. Wash twice using themagnetic separation rack with 0.2 M triethanolamine, pH 8.2 as the washing buffer.2.Resuspend the beads in 1 ml of 20 mM dimetyl pimelimidate dihydrochloride (DMP) in 0.2 M triethanolamine, pH 8.2(5.4 mg DMP/ml buffer). This cross‐linking solution must be prepared freshly.3.Incubate the beads with rotational mixing for 30 minutes at room temperature. Use the magnetic separation rack tocollect the beads and discard the supernatant.4.Resuspend the beads in 1 ml of 50 mM Tris, pH 7.5 to stop the reaction and incubate for 15 minutes at roomtemperature with rotational mixing.e the magnetic separation rack to collect the beads and discard the supernatant.6.Wash the cross‐linked beads three times with 1 ml PBS, pH7.4.2.4.2Binding Antigen to the IgG Cross‐linked Beads1.Add sample containing target antigen to the beads. For a 100 kD protein, use a volume containing approximate 25 µgtarget antigen/ml beads to assure an excess of antigen. If dilution of antigen is necessary, PBS or 0.1 M phosphate buffer (pH 7‐8) can be used as dilution buffer.2.Incubate with tilting and rotation for one hour at room temperature.3.Place the tube on the magnetic separation rack for 2 minutes to collect the IgG‐coated Beads‐target complex. Forviscous samples, double the time on the rack. Discard the supernatant.4.Wash the beads 3 times using 1 ml PBS.2.4.3Elution of Target ProteinA.Denaturing elution1.Place the tube from section2.4.2 on the magnetic separation rack to collect the beads and discard the supernatant.2.Add 100 µl 1XSDS Sample Buffer to the tube and mix well.3.Heat the tube at 100°C for five minutes.e the magnetic separation rack to collect the beads and transfer the supernatant containing desired sample into anew tube.5.Analyze the sample by SDS‐PAGE followed by Western blot analysis.B.Non‐denaturing elution1.Place the tube from section2.4.2 on the magnetic separation rack to collect the beads and discard the supernatant.2.Add 100 µl Elution Buffer to the tube and mix well. Incubate for five minutes at room temperature with occasionalmixing.e the magnetic separation rack to collect the beads and transfer the supernatant into a new tube.4.Repeat Step 2 and 3 twice.5.Add 10 μl Neutralization Buffer to each 100 μl of eluate to neutralize the pH.3.TroubleshootingReview the information below to troubleshoot your experiments using the GenScript Protein A/G MagBeads. Problem Possible Cause SolutionThe beads are difficult toimmobilize using the magneticseparation rack.Too many beads are used.Decrease the volume of MagBeadssuspension.A considerable amount of samplehas been added, but very fewspecific antibody of interest isdetected.The antibody of interest is at verylow concentration.Use a serum‐free medium for cellsupernatant samples.Affinity‐purify the antibody using itsspecific antigen coupled to anaffinity supporting material.The antibody of interest is purified,but it is degraded (as determined byloss of function in downstreamassay).The antibody is sensitive to low‐pHelution buffer.The downstream application issensitive to the neutralized elutionbuffer.Try another elution reagent, such as3.5 M MgCl2, 10 mM phosphate, pH7.2.Desalt or dialyze the eluted sampleinto a suitable buffer.No antibody is detected in anyeluate.The antibody in the sample cannotbind to Protein A/G.Try GenScript Protein A MagBeadsor Protein G MagBeads.4.General Information4.1Storage and StabilityThis product is stable until the expiration date stated on the COA, when stored unopened at 2–8°C. Do not freeze the product. Keep the MagBeads in liquid suspension during storage and all handling steps. Drying will cause loss of binding capacity and result in reduced performance. Resuspend the beads well before use. Be careful to avoid bacterial/fungal contamination.4.2Technical SupportPlease contact GenScript for further technical information (see contact details). Certificate of Analysis/Compliance is available upon request. The latest revision of the package insert/instructions for use is available on .4.3Warning and LimitationsThis product is for research use only. Not intended for any animal or human therapeutic or diagnostic use unless otherwise stated. This product contains 20 % EtOH as a preservative. Flammable liquid and vapor. Flash point 38°C. R‐10 flammable. Material Safety Data Sheet (MSDS) is available at .4.4Related MagBeads ProductsCat. No. Product NameL00273 Protein A MagBeadsL00274 Protein G MagBeadsL00295 Ni‐Charged MagBeadsL00327 Glutathione MagBeadsL00275 Mouse Anti‐His mAb MagBeadsL00336 Mouse Anti‐GST mAb MagBeadsGenScript USA Inc.860 Centennial Ave.,Piscataway, NJ 08854Toll‐Free: 1‐877‐436‐7274Tel: 1‐732‐885‐9188, Fax: 1‐732‐210‐0262Email: *********************Web: 。
巴斯夫露保康
安全技术说明书页: 1/14 巴斯夫安全技术说明书按照GB/T 16483编制日期 / 本次修订: 07.11.2022版本: 7.0日期/上次修订: 18.12.2021上次版本: 6.1日期 / 首次编制: 29.12.2005产品: 露保康®Product: Lupro-Grain®(30062123/SDS_GEN_CN/ZH)印刷日期 10.09.20231. 化学品及企业标识露保康®Lupro-Grain®推荐用途和限制用途: 饲料添加剂公司:巴斯夫(中国)有限公司中国上海浦东江心沙路300号邮政编码 200137电话: +86 21 20391000传真号: +86 21 20394800E-mail地址: **********************紧急联络信息:巴斯夫紧急热线中心（中国）+86 21 5861-1199巴斯夫紧急热线中心（国际）:电话: +49 180 2273-112Company:BASF (China) Co., Ltd.300 Jiang Xin Sha RoadPu Dong Shanghai 200137, CHINA Telephone: +86 21 20391000Telefax number: +86 21 20394800E-mail address: ********************** Emergency information:Emergency Call Center (China):+86 21 5861-1199International emergency number: Telephone: +49 180 2273-1122. 危险性概述纯物质和混合物的分类:皮肤腐蚀/刺激: 分类2巴斯夫安全技术说明书日期 / 本次修订: 07.11.2022版本: 7.0产品: 露保康®Product: Lupro-Grain®(30062123/SDS_GEN_CN/ZH)印刷日期 10.09.2023易燃液体: 分类3急性毒性: 分类5 (口服)急性毒性: 分类5 (皮肤接触)严重损伤/刺激眼睛: 分类1特异性靶器官毒性-一次接触: 分类3 (对呼吸道系统有刺激性)标签要素和警示性说明:图形符号警示词:危险危险性说明:H226易燃液体和蒸气。
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Anti-DYKDDDDK Magarose Beads
目录
1.产品介绍 (1)
2.试剂准备 (1)
3.样品纯化 (2)
4.试剂兼容性 (3)
5.问题及解决方案 (3)
6.订购信息及相关产品 (4)
1.产品介绍
Flag标签是一个由八个亲水氨基酸组成的多肽片段，定位在融合蛋白表面，因此更易与抗体结合以及被肠激酶分解。
Magarose Beads系列产品具有超顺磁性、快速磁响应性、丰富羟基官能团和相对集中的粒径等特点，是医学与分子生物学研究中重要的载体工具。
Anti-DYKDDDDK Magarose Beads是以抗flag(DYKDDDDK)抗体为亲和配体，一步纯化原核、酵母或哺乳动物细胞表达的flag标签融合蛋白。
表1.Anti-DYKDDDDK Magarose Beads产品性能
性能指标
基质磁性琼脂糖微球
配体Anti-DYKDDDDK Antibody
结合能力(/ml磁珠)>1mg DYKDDDDK标签蛋白
粒径(μm)30-100
储存缓冲液PBS，0.01%Tween-20,0.02%NaN3
磁珠体积磁珠体积占悬浮液体积的20%
储存温度2°C-8°C
2.试剂准备
2.1样品准备
上柱之前要确保样品溶液有合适的离子强度和pH值，可以用平衡液对样品或细胞培养液稀释，或者用平衡液透析。
样品在上样前建议离心或用0.22μm或0.45μm滤膜过滤，减少杂质，提高蛋白纯化效率和防止堵塞柱子。
2.2缓冲液的准备
所用水和缓冲液在使用之前建议用0.22μm或0.45μm滤膜过滤。
平衡/洗杂液:50mM Tris,0.15M NaCl,pH7.4
洗脱液:0.1M glycine HCl,pH3.0
竞争性洗脱液：50mM Tris,0.15M NaCl,100-500ug flag多肽/ml，pH7.4
中和液：1M Tris-HCl，pH8.0
3.样品纯化
3.1磁珠预处理
将Anti-DYKDDDDK Magarose Beads颠倒数次，保证磁珠完全混匀，取计算量的磁珠悬浮液，转移至离心管中，放置在磁分离器上，静置大约1min，待溶液变澄清后，用移液器吸弃清液。
将离心管磁分离器上取下来，加入与悬浮液等体积的平衡液，使用枪头反复吹打5次，将离心管置于磁分离器上，大约1min，待溶液变澄清后，用移液器吸弃清液，重复洗涤2次。
3.2样品吸附
在步骤3.1预处理的磁珠管中加入样品溶液，漩涡振荡均匀，在室温下置于翻转混合仪或者手工轻轻翻转离心管，促使样品和磁珠充分接触并吸附，约30min后，置于磁分离器上，大约1min，待溶液变澄清后，吸弃上清液。
3.3洗杂
向离心管中加入5倍磁珠体积的洗杂液，振荡悬浮，置于磁分离器上，大约1min，待溶液变澄清后，吸弃上清液。
该操作重复两次以上。
3.4样品洗脱
A酸性洗脱：
在上述离心管中加入3-5倍磁珠体积的酸性洗脱液（0.1M glycine HCl,pH3.0），用移液器吹打5次，然后在室温下置于翻转混合仪或者手工轻轻翻转离心管，5-10min后，置于磁分离器上，大约1min，待溶液变澄清后，吸取上清液，收集洗脱组分，即为目标抗体。
该操作可重复一次。
向洗脱组分中加入洗脱体积十分之一的中和液，调节pH值至7.0-8.0。
注：酸性洗脱后磁珠要立即用平衡液平衡，Anti-DYKDDDDK Magarose Beads在洗脱液中不要超过20min。
B竞争性洗脱：
使用3-5倍磁珠体积的竞争性洗脱液flag多肽100µg/ml洗脱，2-8度孵育30min,将离心管置于磁分离器上，大约1min，待溶液变澄清后，小心取出上清，不要吸到填料。
洗脱样品放置4度，长时间放置-20度保存。
C:变性洗脱
样品缓冲液中含有β-巯基乙醇和DTT,可以使填料中抗体重链和轻链断开（50和25KD）。
含有SDS的样品缓冲液可以使Anti-Flag抗体变性，洗脱后的Anti-DYKDDDDK Magarose Beads没办法重复使用。
每管中加入磁珠体积等量的2X样品缓冲液，煮3min。
将离心管置于磁分离器上，大约1min，
待溶液变澄清后，吸取上清SDS-PAGE电泳检测。
注：Anti-DYKDDDDK Magarose Beads变性洗脱后不能重复使用。
4．试剂兼容性
表2、Anti-DYKDDDDK Magarose Beads试剂兼容性
试剂名称最大耐受浓度备注
β-巯基乙醇10mM纯化过程中应避免使用，如果在IP中
使用，填料不能回收重复利用。
DTT80mM
SDS--
EDTA5mM过高的EDTA会降低蛋白回收率。
Tween-205%过高浓度会影响标签蛋白结合效率。
Triton X-1005%
NP-404%
盐酸胍0.3M过高浓度会使抗体变性。
尿素 1.5M
甘油20%过高浓度会影响标签蛋白结合。
氯化钠1M减少非特异性吸附
5.问题及解决方案
问题原因分析推荐解决方案
流穿中有目的蛋白过载减少上样体积或增加磁珠体积。
结合时间太短延长样品和磁珠的结合时间。
标签未暴露可以加入低浓度的变性剂，上样前透析。
溶液中试剂不兼容样品上样前进行透析。
洗脱组分中没有目的蛋白目的蛋白不稳定使用新配制的样品。
低温操作。
细胞裂解液中加入蛋白酶抑制剂。
样品中无标签融合蛋白纯化前Western检测是否有flag标签融合蛋白目的蛋白表达量太低优化蛋白表达量。
增加上样量。
减少NaCl浓度。
背景太杂非特异性吸附减少上样量
洗杂不充分增加洗杂次数，每次清洗孵育5-10min。
增加洗杂液中盐离子浓度。
6.订购信息及相关产品
名称货号规格
Anti-DYKDDDDK Magarose Beads SM009011ml SM009055ml
Anti-DYKDDDDK Affinity Beads SA0420011ml
SA0420025ml。